By Bob Axmear

This is an overall description of tissue culturing hostas. It is meant to give you a rough idea of how to go about it. You might need to do a little more research on your own. I am going to add to it as things happen, how to remove the explants, etc when they come up. Also Carol Stiff is very helpful and conducts seminars and sells tc products in small quantities. Also a good place for much of the supplies is PhytoTechnology Laboratories, you can buy prepared media there if you want to go that route. A good beginner book is Plants from Test Tubes. Some TC lists are here. Some CDs, I found Carol's informative for beginners, Dr. Ronald A. de Fossard is for more advanced people.

More videos

At this time I am going to show you 3 plants I treated with Bap 10. This is the hormone that causes dormant buds to break open and start growing, these are what you use to collect your explants(to remove living tissue especially to a medium for tissue culture). I treated these three plants on 5-26-2007. I used the hormone at a 3% solution. This means 30ml per liter or 1 oz in a quart of water or fractions of these amounts for making small amounts of solution. Also add a drop or two of surfactant, dish soap works fine. I just use a spray bottle and apply to the leaves and especially to the crown of the plant, down between the leaves. Within about 10 days you should see all kinds of new shoots coming up at the base of the plants.
I want to caution people who want to do this just for more divisions(using bap 10), many times you get a huge increase in divisions but they and the plant stay much smaller for a long period of time, a year or two isn't unusal. Also if you use it for that water and fertilize more to help the plant with all the new divisions.

bap 10

Treated Plants

Within about 10 days you should see all kinds of new shoots just starting to poke out of the soil.

Treated Plants without shoots yet

Dormant Buds for  harvesting explants

Many buds to remove for their mersistem

Dormant Buds for  harvesting explants

Bud removed from the plant with a little of the base tissue. I use a hooked scalpel to do this but whatever works for you.

Hooked scalpel

Bud removed from plant

You need to clean the bud of the outside leaf tissue down to the remaining meristem and a little of the base, it peels like an onion. Make sure it is as clean and white as you can get it by trimming any discoloration off of the base tissue. That is easy to do, just shave the sides of the base with your scalpel.

Bud cleaned down to meristem

In the coming days I will add more photos and a general media recipe or protocol for hostas, this will be for initiation and multiplication of the explants. You can also find recipes in the search engines for this. I will add a basic list of tools and equipment you will need as well soon.


Tools and Equipment Needed
Here is a list of links you can also find supplies as well as Kitchen Culture Kits website

1000 ml beaker or graduated cylinder
Ebay
Ebay
PhyTo Tech

Scalpel and blades, buy assortment of blade styles, hooked ones seem to work best for removing explant. Make sure blade fits handle size as well
Ebay
Phyto Tech

6" or 8" Forceps
Ebay
Kitchen Culture Kit
Phyto Tech

Scale
Ebay
Google

8oz Baby Food Jars, can be used or new, these can be purchased in varying amounts
PhyTo Tech
Ebay

Lids or Closures, also known as Magenta B Caps
Phyto
Google

PH Paper or Meter
Google

Pressure Cooker, found 8 quart works best
Ebay

Pipettes 1 ml for sure, 5 ml comes in handy as well
Ebay

Clear Plastic Bags to put jars in, nice if large enough for nursery tray but could be smaller for smaller amounts of jars
Ebay
Google


List of Chemicals
Agar, only easy to order source I could find
Common White Granulated Sugar
Murashige and Skoog Basal Medium with Gamborg Vitamins, get the 1 Liter packets
BA Benzyladenine
Potassium Phosphate
NAA Naphtaleneacetic Acid Solution



Formulas or Protocols

Take 1 to 4 The protocol isn't too complicated and if someone wants to continue on I can sure explain it easily enough. Here is a link to one so you can see it. Also here is a link to some videos online.   Another

This way if someone wants to do it just email me and I can explain it further or call, 563 568-4859. Easier to explain than to write it down. Tissue culture isn't that hard, just seems overwhelming. Taken one step at a time the first time or two until you get the hang of it. Don't let the metric system frighten you either, not a problem. The hardest part is keeping everything sterile, the rest is actually pretty easy. Once you learn it you can do all kinds of plants like orchid seed and african violets, etc.

I plan on adding more but just never seem to get around to finishing this. Maybe this winter I can add more details and photos. Basically you just measure the 5 chemicals/hormones and mix them together and sterilize them in baby food jars in the microwave or pressure cooker. Then sterilize the piece of plant with dilute chlorox and then insert it into the chemicals/hormones. Then under the lights they go.


Homemade Box

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